New immunogenic compositions

Abstract

The present invention relates to an immunogenic composition for Proteobacteria protection and reduced transmission. We have identified Proteobacteria serovar variant combinations that generate an immune response capable of robustly driving bacterial enteropathogens into an evolutionary dead end and reducing the transmission of the bacterium. These inactivated immunogenic positions and typically oral vaccines are easy to apply, cheap to produce, and can be stored long-term without cold-chain requirements making them ideal for application in livestock, or in resource-poor areas. They are believed to be the only immunogenic compositions and vaccine formulations capable of breaking the chain of transmission for these types of pathogen.

1 . An immunogenic composition comprising at least two or more inactivated variant serovar of a Proteobacteria strain, wherein each of said two or more inactivated variant serovar comprises a genetic modification of the O-antigen modifying genes, wherein independently each of said genetic modification comprises a glucosylation and/or O-acetylation of the O-antigen. 
     
     
         2 . The immunogenic composition according to  claim 1 , wherein the composition additionally comprises the inactivated wild type serovar of said Proteobacteria strain. 
     
     
         3 . An immunogenic composition according to  claim 1  or  2 , wherein the genetic modification of the O-antigen modifying genes are created following deletions and/or epigenetic modifications of the O-antigen modifying genes Abequose O-acetyl transferase (OafA gene) and gtrABC operons (gtrC gene). 
     
     
         4 . The immunogenic composition according to  claims 1  to  3 , wherein the immunogenic composition induces in an animal, upon infection with the wild type serovar of said Proteobacteria, the production of a serovar of said Proteobacteria strain with a decreased virulence, compared to animals not exposed to the immunogenic composition, wherein decreased virulence is measured as a decrease in mortality and/or morbidity of the infected animal. 
     
     
         5 . The immunogenic composition according to any one of  claims 1 - 4 , wherein the immunogenic composition induces in an animal, upon infection with the wild type serovar of said Proteobacteria, the production of a serovar of said Proteobacteria strain with a decreased transmission rate, compared to animals not exposed to the immunogenic composition. 
     
     
         6 . The immunogenic composition according to anyone of  claims 1 - 5 , wherein the Proteobacteria is  Salmonella  spp or  Escherichia coli , preferably wherein the Proteobacteria is  Salmonella enterica.    
     
     
         7 . The immunogenic composition according to  claim 6 , wherein the  Salmonella enterica  is  Salmonella enterica  subspecies  enterica  serovar  Typhimurium.    
     
     
         8 . The immunogenic composition according to  claim 7 , wherein the  Salmonella enterica  subspecies  enterica Typhimurium  is serovar  Typhimurium  strain ATCC SL1344 or ATCC 14028. 
     
     
         9 . The immunogenic composition according to any one of  claims 1 - 8 , wherein the Proteobacteria is  Salmonella enterica  serovar  Typhimurium , and wherein said at least 2 inactivated serovar of said Proteobacteria strain are selected from O:4[5],12-0 serovar variant, O:4, 12-0 serovar variant, O:4[5],12-2 serovar variant and O:4,12-2 serovar variant. 
     
     
         10 . The immunogenic composition according to any one of  claims 1 - 9 , wherein the Proteobacteria is  Salmonella enterica  subspecies  enterica  serovar  Typhimurium , and wherein said composition comprises at least 4, preferably exactly 4, inactivated serovar variants of said Proteobacteria strain, and wherein said 4 inactivated serovar variants are O:4[5],12-0 serovar variant, O:4, 12-0 serovar variant, O:4[5],12-2 serovar variant and O:4,12-2 serovar variant, and wherein preferably the O-antigen of said four inactivated serovar variants comprises a glycan structure selected from the following formula: 
       
         
           
           
               
               
           
         
       
     
     
         11 . The immunogenic composition according to anyone of  claims 1 - 10  for use as a prophylactic treatment against a disease caused by Proteobacteria in a subject, wherein preferably the subject following treatment contains only a non-transmissible form of said Proteobacteria. 
     
     
         12 . The immunogenic composition according to anyone of  claims 1 - 10  for use in a method of inducing an immune response against a disease caused by Proteobacteria in a subject, wherein said method comprises administrating said immunogenic composition to said subject in need thereof, and wherein preferably administration is by intranasal, intramuscular, subcutaneous, transdermal or sublingual administration. 
     
     
         13 . The immunogenic composition for use according to  claim 11  or  12 , wherein said subject is selected from a human and an agricultural animal, preferably wherein said agricultural animal is selected from the group consisting of: cattle, poultry, swine, horses, sheep and goats. 
     
     
         14 . An immunogenic composition according to anyone of  claims 1 - 10 , for use in preventing a disease caused by Proteobacteria selected from bacterial gastroenteritis, bacterial enterocolitis, urinary tract infection, mastitis, bacterial pneumonia and bacterial sepsis. 
     
     
         15 . A method of generating an immunogenic composition according to any one of  claims 1 - 10 , comprising the following steps:
 i. Administer an inactivated wild type Proteobacterial strain to a subject,   ii. Challenge the subject with the wildtype Proteobacterial strain,   iii. Isolate clones of said Proteobacterial strain from the subject between 2 hours to 7 days post infection,   iv. Identify isolated clones with reduced binding affinity to the wild type Proteobacteria anti-O-antigen antibody,   v. Generate recombinant clones identical to said isolated clones with reduced binding affinity,   vi. Inactivate and combine the recombinant clones with the inactivated wild type Proteobacterial strain,   vii. Repeat steps i-viii, administering the inactivated combined recombinant clones with the inactivated wild-type Proteobacterial strain,   viii. Combine all identified inactivated clones with the inactivated wild type Proteobacteria to produce the immunogenic composition.